1. Field of the Invention
The present invention pertains to the fields of biomanufacturing and infra-red spectroscopy.
2. Related Art
Biomaterial is used as component material in biomanufacturing. For example, biomaterial is a component material in biologically active pharmaceutical ingredient (BAPI) manufacture. Such biomaterial can include, but is not limited to, proteins, DNA fragments, cDNA, and messenger RNA. Biomaterial is generally made up of one or more biomolecules.
Quality monitoring and control are especially important in biomanufacturing. One conventional approach to quality control is to view the biomanufacturing process as a xe2x80x9cblack box.xe2x80x9d Clinical tests and trials are performed on subjects using the final biomanufactured product (i.e., a pharmaceutical). Such clinical testing can be costly and subject to rigorous Federal Drug Administration regulation.
Off-line sampling and analytical techniques are also used to monitor and control processes in biomanufacturing. Infra-red spectroscopy, including FTIR, is used to study samples drawn from a biomanufacturing process off-line. Drawing such samples, however, can be invasive to a biomanufacturing process creating sterilization and other problems. Off-line sampling also has limited value in the quality control of commercial biomanufacturing. Such off-line analysis can involve time-consuming analyte or sample preparation and may only approximate in situ conditions in a stage of biomanufacture. Off-line sampling is too slow for practical commercial biomanufacturing, such as, BAPI manufacture. Data results even from an IR spectrometer are obtained after too long a delay to provide an adequate quality control response to a biomanufacturing stage.
Currently available processes for measuring the stability of biomolecules in bulk storage are tedious, expensive, and time consuming. According to current practice, accelerated stability studies are used to determine the xe2x80x9cshelf lifexe2x80x9d of a biomolecule formulation. These studies involve storage at elevated temperatures, and analysis of the stability of the biomolecule over time, done by sampling techniques. These results are then mathematically fitted to lower temperatures. Although these accelerated studies are allowed by the Food and Drug Administration for pharmaceuticals, the limits are very narrow, because the error can be significant. Since the error in these studies can be high, manufacture of biomolecule formulations requires the skilled artisan to include a certain amount of overage in any given formulation, to account for any unknown amount of degradation. This can be very costly.
Infrared spectroscopy (IR) has long been used in the evaluation of chemical compounds. Fourier Transform Infrared Spectroscopy (FTIR) has been used to identify and evaluate organic and inorganic materials or compounds. See, e.g., Smith, B., Fundamentals of Fourier Transform Infrared Spectroscopy, CRC Press (1996), which is incorporated herein by reference. Using FTIR, spectral data is collected and converted from an interference pattern to a spectrum. The system provides for subtractive elimination of background spectra, such that particular chemical compounds can be identified by a molecular xe2x80x9cfingerprint.xe2x80x9d Organic compounds have been studied using IR spectroscopy, including FTIR spectroscopy, in off-line sampling or analytic applications, but not as a real time method of monitoring and controlling the course of a bioprocess in commercial biomanufacturing.
As recognized by the inventors, what is needed is a method and system for providing real-time, in situ monitoring and control for a complete biomanufacturing process. A biomolecule and its production needs to be monitored and appropriately characterized for the product""s stage of development, in situ and in real-time in different stages of a commercial biomanufacturing process. See, e.g., Faulkner, J., BioPharm, June 2000:26-34, the disclosure of which is incorporated herein by reference in its entirety. Control strategies in response to real-time IR spectroscopic data are needed in each stage of a biomanufacturing process.
The present invention provides real-time, biomanufacturing process monitoring and control in response to infra-red (IR) spectroscopic monitoring of the biomanufacturing process, and fingerprinting of a biomolecule. IR spectroscopy data is used to provide optimal production control for a biomolecule process and is then used to fingerprint the biomolecule in situ in a biomanufacturing process. In one embodiment, Fourier transform infrared spectroscopy (FTIR) is used to monitor the production of a biomolecule and to fingerprint, both qualitatively and quantitatively, the biomolecule at different stages of a biomanufacturing process. In one example, such FTIR fingerprinting is used to differentiate, in real time, between an active or a non-active biomolecule during the stages of a biomanufacturing process, and to control the biomanufacturing process through feedback inputs to optimize the yield of the active form of the biomolecule.
In one preferred embodiment, the biomanufacturing process manufactures a biomaterial in bulk. The biomanufacturing process has four stages: bioproduction (e.g., fermentation), recovery, purification, and bulk formulation and storage. In the bioproduction stage, IR spectroscopy is used (a) to monitor and control homeostasis of the bioproduction reaction, thereby maintaining optimal conditions for increase in biomass and biomolecule synthesis, (b) in some embodiments requiring a two-step growth and induction process, to determine or alternatively detect the optimal time to induce biomolecule synthesis, (c) in some embodiments where the biomolecule is in solution during bioproduction, to monitor, in real-time, in situ, the proportion of the pharmacologically active form of a biomolecule relative to inactive forms, (d) to periodically or continuously adjust the conditions of the biomanufacturing process in order to preferentially favor or alternatively optimize the yield of the pharmacologically active form of the biomolecule, and/or (e) any full or partial combination of (a) through (d). In the recovery and purification stages, IR spectroscopy is used to monitor, in real-time, in situ, the proportion of the pharmacologically active form of a biomolecule relative to inactive forms, and to periodically or continuously adjust the conditions of the biomanufacturing process in order to optimize the yield of the pharmacologically active form of the biomolecule. The presence of an appropriately-characterized biomolecule is verified in situ and in real-time in different stages of a commercial biomanufacturing process. In the bulk storage stage, IR spectroscopy is used to continuously monitor the quality of the stored biomolecule to precisely determine the pharmacological activity of the formulation when it is processed for final finish and fill, and to provide immediate feedback adjustments in the storage conditions to optimize and extend storage. A real-time stability curve during bulk storage will also allow for accurate prediction or extrapolation of the stability of the formulation after finish and fill, thereby minimizing the need for overage. Real-time IR monitoring of bulk storage also provides the potential to automate and accelerate product stability determinations.
Control strategies in response to real-time IR spectroscopic data are provided in each stage of BAPI manufacturing. IR analysis is provided in situ, in real-time to control a bioproduction stage, one or more steps of a recovery stage, one or more steps of a purification stage and/or a bulk storage stage. In this way, both production and development time and costs are minimized.
In addition, biomanufacturing process monitoring and control in response to infra-red (IR) spectroscopic monitoring of the biomanufacturing process, and fingerprinting of a biomolecule can be used to ensure consistency in biomanufacturing processes carried out in different biomanufacturing plants. This advantage of the present invention provides flexibility to the manufacturer to outsource production, and thereby more efficiently control production without sacrificing quality control.
Further embodiments, features, and advantages of the present inventions, as well as the structure and operation of the various embodiments of the present invention, are described in detail below with reference to the accompanying drawings.